Detection of hepatitis "C" virus in formalin-fixed liver tissue by nested polymerase chain reaction

J Med Virol. 1992 Aug;37(4):310-4. doi: 10.1002/jmv.1890370415.

Abstract

Interpretation of antibody to hepatitis C virus (HCV) in patients with liver disease is difficult due to false-positive reactivity in some conditions. To evaluate the feasibility of HCV in archival material, HCV was sought in formalin-fixed, paraffin-embedded liver biopsy specimens. Nested polymerase chain reaction was used to detect hepatitis C virus in formalin-fixed, paraffin-embedded liver biopsy specimens after total RNA was extracted from tissue by proteinase K digestion and phenol/chloroform purification. The relative efficiency of amplification of HCV RNA from formalin-fixed material was estimated semiquantitatively by serial dilution of cDNA synthesised from RNA extracted from fresh and formalin-fixed sections from the same liver. Although HCV RNA could be detected in formalin-fixed liver tissue by nested PCR in 5/5 cases in which HCV was detected in serum, amplification was approximately 5-fold less efficient than when HCV was amplified from fresh tissue. Nevertheless, nested PCR of HCV from formalin-fixed liver tissue represents a useful technique in addressing some important questions related to the pathogenesis of liver disease.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adult
  • Base Sequence
  • Female
  • Formaldehyde
  • Hepacivirus / genetics
  • Hepacivirus / isolation & purification*
  • Hepatitis C / diagnosis
  • Humans
  • Liver / microbiology*
  • Male
  • Middle Aged
  • Molecular Sequence Data
  • Polymerase Chain Reaction / methods*
  • RNA, Viral / genetics
  • Sensitivity and Specificity
  • Tissue Fixation

Substances

  • RNA, Viral
  • Formaldehyde